Daniel:Notebook/HiResChrPaint/2013-5-22: Difference between revisions

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  7. Incubate in the dark for 1 hour
  7. Incubate in the dark for 1 hour
  8. Add 40 uL nf H20
  8. Add 40 uL nf H20
  9. [[Daniel:Protocols/Qiaquick_Column|Column purification]], add 10x PB, elute with 30 uL EB
  9. [[Daniel:Protocols/Qiaquick_Column|Column purification]], add 10x PB, elute with 30 uL nfH20
  10. Measure in nanodrop using microarray setting
  10. Measure in nanodrop using microarray setting
===Nanodrop Results===
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext>
|- style="font-size:12pt;font-weight:bold" align="center" valign="bottom"
| width="65" height="15" | ng/uL DNA
| width="65" | pmol/uL dye
| width="65" | pmol/uL DNA
| width="65" | dye/probe
| width="65" | ug dye
| width="65" | Yield (%)
|- style="font-size:12pt" align="center" valign="bottom"
| align="center" height="15" | 16.5
| align="center" | 2.2
| align="center" | 0.62
| align="center" | 3.564
| align="center" | 0.495
| align="center" | 11.0
|}
===Discussion===

Latest revision as of 20:52, 22 May 2013

Probe Prep Test 2 (Started 5/10/2013)[edit]

Back to Calendar

Dye Coupling (Round 2)[edit]

After talking with Dr. Zhang yesterday, he suggested I try the dye coupling with just 1 reaction, and purify with a standard column purification.

Protocol[edit]

1. Warm DMSO, Sodium bicarbonate, and V4S3 in 37 degree shaker
2. Add 2.5 uL nf H20 to dna (should bring total to ~5)
3. Denature at 95C for 5 minutes
4. Add 3 uL sodium bicarb to reaction
5. Add 2 uL DMSO to dye, vortex ≥10 seconds
6. Add 2 uL dye/DMSO to reaction; vortex ≥10 seconds
7. Incubate in the dark for 1 hour
8. Add 40 uL nf H20
9. Column purification, add 10x PB, elute with 30 uL nfH20
10. Measure in nanodrop using microarray setting

Nanodrop Results[edit]

ng/uL DNA pmol/uL dye pmol/uL DNA dye/probe ug dye Yield (%)
16.5 2.2 0.62 3.564 0.495 11.0

Discussion[edit]